ICS 07.080 CCS Z51 团 体 标 准 T/ SSESB 14 -2025 基于环境 DNA技术的水生入侵物种多重 PCR 检测方法 Multiplex PCR detection method for aquatic invasive species based on environmental DNA technology 2025 -04-25 发布 2025 -04-25 实施 上海市环境科学学会 发布 全国团体标准信息平台 全国团体标准信息平台 T/ SSESB 14 -2025 i 目 次 前 言 ................................ ................................ ................................ ................................ ii 1 范围 ................................ ................................ ................................ ................................ 1 2 规范性引用文件 ................................ ................................ ................................ ............ 1 3 术语和定义 ................................ ................................ ................................ .................... 1 4 技术原理 ................................ ................................ ................................ ........................ 2 5 引物 ................................ ................................ ................................ ................................ 3 6 试剂 ................................ ................................ ................................ ................................ 3 6.1 采样及前处理试剂 ................................ ................................ ............................... 3 6.2 环境 DNA提取试剂 ................................ ................................ ............................ 4 6.3 PCR扩增试剂 ................................ ................................ ................................ ....... 4 6.4 凝胶电泳试剂 ................................ ................................ ................................ ....... 4 7 仪器设备 ................................ ................................ ................................ ........................ 4 8 检测步骤 ................................ ................................ ................................ ........................ 5 8.1 水样采集 ................................ ................................ ................................ ............... 5 8.2 水样过滤 ................................ ................................ ................................ ............... 5 8.3 DNA 提取 ................................ ................................ ................................ .............. 5 8.4 DNA 质量检测和保存 ................................ ................................ .......................... 5 8.5 多重 PCR扩增 ................................ ................................ ................................ ..... 6 8.6 凝胶电泳检测 ................................ ................................ ................................ ....... 6 8.7 PCR产物纯化和保存 ................................ ................................ ........................... 6 8.8 测序分析 ................................ ................................ ................................ ............... 6 9 结果判定 ................................ ................................ ................................ ........................ 7 10 质量控制与保证 ................................ ................................ ................................ .......... 7 10.1 试验对照设置 ................................ ................................ ................................ ..... 7 10.2 平行样品设置 ................................ ................................ ................................ ..... 7 10.3 质量保证 ................................ ................................ ................................ ............. 7 11 废弃物处理 ................................ ................................ ................................ .................. 7 附 录 A（资料性附录） 外来入侵物种基因扩增靶标序列 ................................ ........ 8 全国团体标准信息平台 T/ SSESB 14 -2025 ii